The aim of this project is to culture neurons in the laboratory for use in studies of axoplasmic structure and transport. These cultured neurons are also to be used in connection with voltage clamp and patch clamp experiments of ionic channels and their conductances and gating mechanisms. A tissue culture (TC) laboratory has been established. A sterile tissue culture hood, CO2 incubator, refrigerators and freezer all restricted to TC use in augmented by U.V. germicidal lights and sterile techniques. Plastic TC flasks, dishes, pipettes, filters, etc. are essential because of lack of proper TC washing facilities. Autoclaves and a sterilizing oven are available. Squid embryos are cultured up to hatching in separate sea water table. Squid fertilized fingers are decontaminated with dilute bleach (0.6%) in artificial sea water (ASW), dissected from the egg jelly and chorions with iridectomy scissors and washed with ASW containing concentrated antibiotics in sterile leucocyte tubes under the hood. Standard TC medium is Hanks minimal essential amino acids and vitamins dissolved in ASW sterilized via 0.22 Mum filters. Fetal bovine serum and chick embryo extract is added as needed for squid embryo cell growth and maintenance. Whole embryos remain alive in the medium for one to two weeks. The culture of neurons is still in the early phase but appears promising.